Journal Article
. 1997 Jul; 159(1):144-51.

Interaction of CTLA-4 with the clathrin-associated protein AP50 results in ligand-independent endocytosis that limits cell surface expression

E Chuang 1 M L Alegre  C S Duckett  P J Noel  M G Vander Heiden  C B Thompson  
Affiliations
  • PMID: 9200449
  •     66 citations

Abstract

CTLA-4 is a lymphocyte cell surface receptor expressed by activated T cells that functions to down-regulate T cell responses induced by TCR and CD28 stimulation. Since CTLA-4 competes with CD28 for binding to the common ligands B7-1 and B7-2, the level of CTLA-4 surface expression is likely to play an important role in its ability to inhibit CD28-dependent T cell activation. The factors that regulate these levels are poorly understood. Recent studies have revealed that following T cell activation, the majority of CTLA-4 is localized intracellularly rather than on the cell surface, and surface CTLA-4 is rapidly reinternalized. In this study, we investigate the molecular mechanism underlying the rapid clearance of CTLA-4 from the cell surface. The data demonstrate that cell surface CTLA-4 is endocytosed into clathrin-coated vesicles even in the absence of ligand. The targeting of CTLA-4 to clathrin-coated vesicles is mediated by the clathrin-associated adaptor complex AP-2. The cytoplasmic domain of CTLA-4 was found to specifically bind to AP50, the medium chain subunit of AP-2 in both yeast two-hybrid and coimmunoprecipitation assays. The interaction requires the peptide sequence 199-GVYVKM-204 in the cytoplasmic tail of CTLA-4. Mutation of the CTLA-4 amino acid residue Y201 abrogates the interaction with AP50, resulting in the accumulation of CTLA-4 at the cell surface. Together these data suggest that the interaction of CTLA-4 with AP50 plays an important role in regulating the cell surface expression of CTLA-4.

Other Links

B7-1 engagement of cytotoxic T lymphocyte antigen 4 inhibits T cell activation in the absence of CD28.
F Fallarino, P E Fields, T F Gajewski.
J Exp Med, 1998 Jul 07; 188(1). PMID: 9653097    Free PMC article.
Role of the CTLA-4 receptor in T cell activation and immunity. Physiologic function of the CTLA-4 receptor.
P Scheipers, H Reiser.
Immunol Res, 1998 Dec 09; 18(2). PMID: 9844829
Review.
Expression of CTLA-4 molecule in peripheral blood T lymphocytes from patients with systemic lupus erythematosus.
M F Liu, H S Liu, C R Wang, H Y Lei.
J Clin Immunol, 1998 Dec 19; 18(6). PMID: 9857283
Defective CTLA-4 cycling pathway in Chediak-Higashi syndrome: a possible mechanism for deregulation of T lymphocyte activation.
F J Barrat, F Le Deist, +4 authors, G de Saint Basile.
Proc Natl Acad Sci U S A, 1999 Jul 21; 96(15). PMID: 10411929    Free PMC article.
Cytotoxic T lymphocyte antigen 4 (CTLA-4) engagement delivers an inhibitory signal through the membrane-proximal region in the absence of the tyrosine motif in the cytoplasmic tail.
C Nakaseko, S Miyatake, +5 authors, T Saito.
J Exp Med, 1999 Sep 28; 190(6). PMID: 10499915    Free PMC article.
Internalization of CD26 by mannose 6-phosphate/insulin-like growth factor II receptor contributes to T cell activation.
H Ikushima, Y Munakata, +5 authors, C Morimoto.
Proc Natl Acad Sci U S A, 2000 Jul 19; 97(15). PMID: 10900005    Free PMC article.
CD28, CTLA-4 and their ligands: who does what and to whom?
D M Sansom.
Immunology, 2000 Sep 30; 101(2). PMID: 11012769    Free PMC article.
Review.
Study of the interaction of the medium chain mu 2 subunit of the clathrin-associated adapter protein complex 2 with cytotoxic T-lymphocyte antigen 4 and CD28.
E R Follows, J C McPheat, +7 authors, W M Abbott.
Biochem J, 2001 Oct 05; 359(Pt 2). PMID: 11583591    Free PMC article.
Cell surface expression of the HIV-1 envelope glycoproteins is directed from intracellular CTLA-4-containing regulated secretory granules.
Luis R Miranda, Brian C Schaefer, +2 authors, Alex Franzusoff.
Proc Natl Acad Sci U S A, 2002 Jun 13; 99(12). PMID: 12060749    Free PMC article.
CTLA-4 and tolerance: the biochemical point of view.
Shunsuke Chikuma, Jeffrey A Bluestone.
Immunol Res, 2004 Jan 10; 28(3). PMID: 14713717
Review.
Multiple functions for CD28 and cytotoxic T lymphocyte antigen-4 during different phases of T cell responses: implications for arthritis and autoimmune diseases.
Monika C Brunner-Weinzierl, Holger Hoff, Gerd-R Burmester.
Arthritis Res Ther, 2004 Apr 03; 6(2). PMID: 15059264    Free PMC article.
Review.
Dileucine and YXXL motifs in the cytoplasmic tail of the bovine leukemia virus transmembrane envelope protein affect protein expression on the cell surface.
Sinisa Novakovic, Earl T Sawai, Kathryn Radke.
J Virol, 2004 Jul 16; 78(15). PMID: 15254202    Free PMC article.
Translocation of CD28 to lipid rafts and costimulation of IL-2.
Ali Sadra, Tomas Cinek, John B Imboden.
Proc Natl Acad Sci U S A, 2004 Jul 29; 101(31). PMID: 15280538    Free PMC article.
Testing for endocytosis in plants.
F Aniento, D G Robinson.
Protoplasma, 2005 Oct 19; 226(1-2). PMID: 16231096
Review.
CTLA-4 interacts with STAT5 and inhibits STAT5-mediated transcription.
M Srahna, L A Van Grunsven, J E Remacle, P Vandenberghe.
Immunology, 2006 Feb 16; 117(3). PMID: 16476059    Free PMC article.
Checkpoint blockade in cancer immunotherapy.
Alan J Korman, Karl S Peggs, James P Allison.
Adv Immunol, 2006 May 30; 90. PMID: 16730267    Free PMC article.
Highly Cited. Review.
Genetic analysis of the exon 1 position 49 CD152 dimorphism in multiple sclerosis.
Rodney Stuart, Amy E Lovett-Racke, +2 authors, Michael K Racke.
J Neuroimmunol, 2007 Oct 09; 191(1-2). PMID: 17920697    Free PMC article.
Review.
Site-specific ubiquitination exposes a linear motif to promote interferon-alpha receptor endocytosis.
K G Suresh Kumar, Hervé Barriere, +8 authors, Serge Y Fuchs.
J Cell Biol, 2007 Dec 07; 179(5). PMID: 18056411    Free PMC article.
CTLA-4 trafficking and surface expression.
Elke Valk, Christopher E Rudd, Helga Schneider.
Trends Immunol, 2008 May 13; 29(6). PMID: 18468488    Free PMC article.
Review.
CTLA-4 is a direct target of Wnt/beta-catenin signaling and is expressed in human melanoma tumors.
Kavita V Shah, Andy J Chien, Cassian Yee, Randall T Moon.
J Invest Dermatol, 2008 Jun 20; 128(12). PMID: 18563180    Free PMC article.
Signals and sequences that control CD28 localization to the central region of the immunological synapse.
Mariano Sanchez-Lockhart, Beth Graf, Jim Miller.
J Immunol, 2008 Nov 20; 181(11). PMID: 19017952    Free PMC article.
CD28 and CTLA-4 coreceptor expression and signal transduction.
Christopher E Rudd, Alison Taylor, Helga Schneider.
Immunol Rev, 2009 May 12; 229(1). PMID: 19426212    Free PMC article.
Highly Cited. Review.
Activation-induced accumulation of B and T lymphocyte attenuator at the immunological synapse in CD4+ T cells.
Takayoshi Owada, Norihiko Watanabe, +7 authors, Hiroshi Nakajima.
J Leukoc Biol, 2009 Nov 07; 87(3). PMID: 19892849    Free PMC article.
Conservation and diversification of dileucine signal recognition by adaptor protein (AP) complex variants.
Rafael Mattera, Markus Boehm, +2 authors, Juan S Bonifacino.
J Biol Chem, 2010 Nov 26; 286(3). PMID: 21097499    Free PMC article.
Trans-endocytosis of CD80 and CD86: a molecular basis for the cell-extrinsic function of CTLA-4.
Omar S Qureshi, Yong Zheng, +12 authors, David M Sansom.
Science, 2011 Apr 09; 332(6029). PMID: 21474713    Free PMC article.
Highly Cited.
Intrinsic and extrinsic control of peripheral T-cell tolerance by costimulatory molecules of the CD28/ B7 family.
Hélène Bour-Jordan, Jonathan H Esensten, +3 authors, Jeffrey A Bluestone.
Immunol Rev, 2011 Apr 15; 241(1). PMID: 21488898    Free PMC article.
Highly Cited. Review.
The emerging role of CTLA4 as a cell-extrinsic regulator of T cell responses.
Lucy S K Walker, David M Sansom.
Nat Rev Immunol, 2011 Nov 26; 11(12). PMID: 22116087
Highly Cited.
Constitutive clathrin-mediated endocytosis of CTLA-4 persists during T cell activation.
Omar S Qureshi, Satdip Kaur, +8 authors, David M Sansom.
J Biol Chem, 2012 Jan 21; 287(12). PMID: 22262842    Free PMC article.
Unmasking the immune recognition of prostate cancer with CTLA4 blockade.
Serena S Kwek, Edward Cha, Lawrence Fong.
Nat Rev Cancer, 2012 Mar 02; 12(4). PMID: 22378189    Free PMC article.
Review.
The N-terminal region of IFITM3 modulates its antiviral activity by regulating IFITM3 cellular localization.
Rui Jia, Qinghua Pan, +5 authors, Chen Liang.
J Virol, 2012 Oct 12; 86(24). PMID: 23055554    Free PMC article.
Highly Cited.
Protein tyrosine phosphatase 1B is a key regulator of IFNAR1 endocytosis and a target for antiviral therapies.
Christopher J Carbone, Hui Zheng, +8 authors, Serge Y Fuchs.
Proc Natl Acad Sci U S A, 2012 Nov 07; 109(47). PMID: 23129613    Free PMC article.
Comparison of the intracellular trafficking itinerary of ctla-4 orthologues.
Satdip Kaur, Omar S Qureshi, David M Sansom.
PLoS One, 2013 Apr 09; 8(4). PMID: 23565286    Free PMC article.
At the bench: preclinical rationale for CTLA-4 and PD-1 blockade as cancer immunotherapy.
Andrew M Intlekofer, Craig B Thompson.
J Leukoc Biol, 2013 Apr 30; 94(1). PMID: 23625198    Free PMC article.
Highly Cited. Review.
ICOS regulates the generation and function of human CD4+ Treg in a CTLA-4 dependent manner.
Jian Zheng, Ping-Lung Chan, +6 authors, Wenwei Tu.
PLoS One, 2013 Dec 07; 8(12). PMID: 24312642    Free PMC article.
Rab8 binding to immune cell-specific adaptor LAX facilitates formation of trans-Golgi network-proximal CTLA-4 vesicles for surface expression.
Matthew C Banton, Kerry L Inder, +2 authors, Helga Schneider.
Mol Cell Biol, 2014 Feb 12; 34(8). PMID: 24515439    Free PMC article.
Murine Tim-1 is excluded from the immunological synapse.
Jean Lin, Leo Chen, Lawrence P Kane.
F1000Res, 2012 Jan 01; 1. PMID: 24627760    Free PMC article.
Tyrosine 201 of the cytoplasmic tail of CTLA-4 critically affects T regulatory cell suppressive function.
Melanie Stumpf, Xuyu Zhou, Shunsuke Chikuma, Jeffrey A Bluestone.
Eur J Immunol, 2014 Mar 22; 44(6). PMID: 24648182    Free PMC article.
The ap-2 clathrin adaptor mediates endocytosis of an inhibitory killer cell Ig-like receptor in human NK cells.
Amanda K Purdy, Diana A Alvarez Arias, +3 authors, Kerry S Campbell.
J Immunol, 2014 Sep 23; 193(9). PMID: 25238755    Free PMC article.
Diverse mechanisms regulate the surface expression of immunotherapeutic target ctla-4.
Helga Schneider, Christopher E Rudd.
Front Immunol, 2014 Dec 30; 5. PMID: 25538704    Free PMC article.
Review.
Confusing signals: recent progress in CTLA-4 biology.
Lucy S K Walker, David M Sansom.
Trends Immunol, 2015 Jan 15; 36(2). PMID: 25582039    Free PMC article.
Highly Cited. Review.
A transendocytosis model of CTLA-4 function predicts its suppressive behavior on regulatory T cells.
Tie Zheng Hou, Omar S Qureshi, +4 authors, David M Sansom.
J Immunol, 2015 Jan 30; 194(5). PMID: 25632005    Free PMC article.
Glutamate Receptor Interacting Protein 1 Regulates CD4(+) CTLA-4 Expression and Transplant Rejection.
K L Modjeski, S C Levy, +5 authors, C N Morrell.
Am J Transplant, 2015 Nov 26; 16(5). PMID: 26601915    Free PMC article.
Cell Type-Specific Regulation of Immunological Synapse Dynamics by B7 Ligand Recognition.
Joanna Brzostek, Nicholas R J Gascoigne, Vasily Rybakin.
Front Immunol, 2016 Feb 13; 7. PMID: 26870040    Free PMC article.
Review.
Acute myeloid leukemia stem cell markers in prognosis and targeted therapy: potential impact of BMI-1, TIM-3 and CLL-1.
Noureldien H E Darwish, Thangirala Sudha, +4 authors, Shaker A Mousa.
Oncotarget, 2016 Aug 11; 7(36). PMID: 27506934    Free PMC article.
Immunometabolic Regulations Mediated by Coinhibitory Receptors and Their Impact on T Cell Immune Responses.
Nikolaos Patsoukis, Jessica D Weaver, +3 authors, Vassiliki A Boussiotis.
Front Immunol, 2017 Apr 27; 8. PMID: 28443090    Free PMC article.
Review.
Immunological effects of hypomethylating agents.
Katherine E Lindblad, Meghali Goswami, Christopher S Hourigan, Karolyn A Oetjen.
Expert Rev Hematol, 2017 Jun 24; 10(8). PMID: 28644756    Free PMC article.
Review.
A reappraisal of CTLA-4 checkpoint blockade in cancer immunotherapy.
Xuexiang Du, Fei Tang, +14 authors, Yang Liu.
Cell Res, 2018 Feb 24; 28(4). PMID: 29472691    Free PMC article.
Highly Cited.
Endocrine Toxicity of Cancer Immunotherapy Targeting Immune Checkpoints.
Lee-Shing Chang, Romualdo Barroso-Sousa, +3 authors, Le Min.
Endocr Rev, 2018 Sep 06; 40(1). PMID: 30184160    Free PMC article.
Review.
Characterization of CTLA4 Trafficking and Implications for Its Function.
Sahamoddin Khailaie, Behzad Rowshanravan, +6 authors, Michael Meyer-Hermann.
Biophys J, 2018 Sep 17; 115(7). PMID: 30219287    Free PMC article.
Human DEF6 deficiency underlies an immunodeficiency syndrome with systemic autoimmunity and aberrant CTLA-4 homeostasis.
Nina K Serwas, Birgit Hoeger, +39 authors, Kaan Boztug.
Nat Commun, 2019 Jul 17; 10(1). PMID: 31308374    Free PMC article.
Molecular thermodynamics of receptor competition for endocytic uptake.
Andre C M DeGroot, Chi Zhao, +2 authors, Jeanne C Stachowiak.
Soft Matter, 2019 Sep 11; 15(37). PMID: 31502617    Free PMC article.
Hijacking antibody-induced CTLA-4 lysosomal degradation for safer and more effective cancer immunotherapy.
Yan Zhang, Xuexiang Du, +9 authors, Yang Liu.
Cell Res, 2019 Jul 04; 29(8). PMID: 31267017    Free PMC article.
The Potential Role of Inhibitory Receptors in the Treatment of Psoriasis.
Neha Shah, Sabina Sandigursky, Adam Mor.
Bull Hosp Jt Dis (2013), 2017 Sep 14; 75(3). PMID: 28902599    Free PMC article.
Review.
CTLA-4: a moving target in immunotherapy.
Behzad Rowshanravan, Neil Halliday, David M Sansom.
Blood, 2017 Nov 10; 131(1). PMID: 29118008    Free PMC article.
Highly Cited. Review.
Leukemic B Cell CTLA-4 Suppresses Costimulation of T Cells.
Priscilla Do, Kyle A Beckwith, +9 authors, John C Byrd.
J Immunol, 2019 Mar 27; 202(9). PMID: 30910862    Free PMC article.
Association of DNA-Methylation Profiles With Immune Responses Elicited in Breast Cancer Patients Immunized With a Carbohydrate-Mimicking Peptide: A Pilot Study.
Cinthia Violeta Hernandez Puente, Ping-Ching Hsu, +8 authors, Behjatolah Monzavi-Karbassi.
Front Oncol, 2020 Jun 26; 10. PMID: 32582547    Free PMC article.
Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells.
Margherita Passariello, Cinzia Vetrei, +7 authors, Claudia De Lorenzo.
Cancers (Basel), 2020 Aug 13; 12(8). PMID: 32781690    Free PMC article.
The role of lysosomes in cancer development and progression.
Tao Tang, Ze-Yu Yang, +7 authors, Shi-Cang Yu.
Cell Biosci, 2020 Dec 10; 10(1). PMID: 33292489    Free PMC article.
Review.
Beyond T-Cells: Functional Characterization of CTLA-4 Expression in Immune and Non-Immune Cell Types.
Damilola Oyewole-Said, Vanaja Konduri, +3 authors, William K Decker.
Front Immunol, 2021 Jan 02; 11. PMID: 33384695    Free PMC article.
Review.
Treg-expressed CTLA-4 depletes CD80/CD86 by trogocytosis, releasing free PD-L1 on antigen-presenting cells.
Murat Tekguc, James Badger Wing, +2 authors, Shimon Sakaguchi.
Proc Natl Acad Sci U S A, 2021 Jul 25; 118(30). PMID: 34301886    Free PMC article.
Regulation of CTLA-4 recycling by LRBA and Rab11.
Daniel Janman, Claudia Hinze, +8 authors, David M Sansom.
Immunology, 2021 May 08; 164(1). PMID: 33960403    Free PMC article.
The efficacy and safety of immunotherapy in thymic epithelial tumors: more effective, more risky: a systematic review.
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J Thorac Dis, 2021 Sep 17; 13(8). PMID: 34527346    Free PMC article.
Review.
Weakly Internalized Receptors Use Coated Vesicle Heterogeneity to Evade Competition during Endocytosis.
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Biochemistry, 2021 Jun 26; 60(27). PMID: 34170686    Free PMC article.
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World J Hepatol, 2021 Oct 12; 13(9). PMID: 34630870    Free PMC article.
Review.
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Tipping the balance: inhibitory checkpoints in intestinal homeostasis.
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Mucosal Immunol, 2018 Dec 01; 12(1). PMID: 30498201
Review.